小麦株高相关基因<i>TaGAMyb-B</i>在水稻中的功能研究

doi:10.7668/hbnxb.20193241

摘要/Abstract

摘要：

为进一步探究小麦的TaGAMyb-B不同等位变异基因对茎秆伸长的作用,利用水稻的农杆菌转化体系、RT-qPCR、组织切片和细胞组织特异性分析等方法系统研究TaGAMyb-B基因不同等位变异中84 bp InDel 的功能。结果发现:在TaGAMyb-B超表达的水稻转基因株系中,该基因在种子、根、茎以及叶中均有表达; 转TaGAMyb-Ba-GFP和TaGAMyb-Bb-GFP基因型水稻T₂种子在应对外界NaCl、GA和甘露醇胁迫处理时,其表现的敏感性为TaGAMyb-Bb-GFP大于TaGAMyb-Ba-GFP;转TaGAMyb-Bb-GFP基因型水稻的第一、二和三茎粗,穗长和分蘖数均显著小于转TaGAMyb-Ba-GFP基因型;转基因水稻后代第二茎间的细胞组织切片分析表明:转TaGAMyb-Ba-GFP基因型水稻横切面厚壁组织细胞的平均厚度显著大于转TaGAMyb-Bb-GFP基因型水稻,并且其厚壁细胞的平均长度极显著小于转TaGAMyb-Bb-GFP基因型水稻。以上结果表明,TaGAMyb-B不同等位变异中84 bp序列缺失在转基因水稻中除了增加非生物胁迫抗性和抗倒伏性功能外,还显著增加了穗长和分蘖数。

关键词: 转基因水稻, TaGAMyb-B, 等位变异, 非生物胁迫, 抗倒伏性

Abstract:

To further investigated the effect of different allelic variants of TaGAMyb-B genes in wheat on stem elongation, used the rice Agrobacterium transformation system,RT-qPCR,tissue section and cell tissue specific analysis to systematically study the function of the 84 bp InDel of TaGAMyb-B.The results showed that,in the over-expressed transgenic rice lines,transcript expression of TaGAMyb-B was detected in seeds,roots,stems and leaves;TaGAMyb-Bb-GFP transgenic seeds were more sensitive under the treatment of NaCl,GA and mannitol compared with TaGAMyb-Ba-GFP;the diameters of first,second and third stem inter-nodes,spike length and tiller number of TaGAMyb-Ba-GFP were significantly larger than that of TaGAMyb-Bb-GFP;and the analysis of cell tissue sections showed that the average thickness of the thick-walled tissue cells in the transverse sections of the transgenic TaGAMyb-Ba-GFP rice was significantly greater than that of the transgenic TaGAMyb-Bb-GFP event,the average length of thick-walled cells was extremely significantly shorter than that of transgenic TaGAMyb-Bb-GFP event.The above mentioned results indicated that the 84 bp deletion in TaGAMyb-B increased not only abiotic stress resistance and plant lodging resistance,but also spike length and tiller number in transgenic rice.

Key words: Transgenic rice, TaGAMyb-B, Allelic variation, Abiotic stress, Lodging resistance

刘娟, 冯玉梅, 韩冰, 邢燕平, 李淑芬, 杨燕. 小麦株高相关基因TaGAMyb-B在水稻中的功能研究[J]. 华北农学报, 2023, 38(3): 42-48. doi: 10.7668/hbnxb.20193241.

LIU Juan, FENG Yumei, HAN Bing, XING Yanping, LI Shufen, YANG Yan. Functional Study of TaGAMyb-B Conferring Wheat Plant Height in Transgenic Rice[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(3): 42-48. doi: 10.7668/hbnxb.20193241.

http://www.hbnxb.net/CN/Y2023/V38/I3/42

图/表 11

表1 研究所用引物序列

Tab.1 The primer sequences used in this study

引物名称 Primer name	上游引物(5'-3') Forward primer(5'-3')	下游引物(5'-3') Reverse primer(5'-3')	退火温度/℃ TM	片段大小/bp Fragment size
GAMyb-B2F/R	CTACAGATTTTTATTTTTATCTATC	CAACAAGAGGTGAACTTTTTT	57.0	659/764
JDF/R	ACTTGTTTGTCGGGTCATCTTTTC	GAAGTAAAGCAGTGAATCAGCGTG	60.0	1 106
Q-OsActinF/R	CCTGACGGAGCGTGGTTAC	CCAGGGCGATGTAGGAAAGC	63.0	321
Q-TaGAMyb-BF/R	GCGAGAATCTTTCCAGTGACCTT	CAATCCAGGGAGTAAATGGTCAGA	60.0	230

图1 重组表达载体酶切鉴定 M.DL10000 bp DNA Marker;1.Ubi-TaGAMyb-Bb-GFP;2.Ubi-TaGAMyb-Ba-GFP.

Fig.1 Identification of the recombinant expression vector by restriction endonucleases

图2 重组表达载体(Ubi-TGAMyb-Ba/b-GFP)PCR鉴定图谱 M.DL1000 bp DNA Marker;1.TaGAMyb-Bb(764 bp);2.TaGAMyb-Ba(659 bp).

Fig.2 Identification of the recombinant expression vector(Ubi-TGAMyb-Ba/b-GFP)by PCR

图3 转基因水稻阳性植株的PCR鉴定 M.DL2000 bp DNA Marker;1-9.部分转基因水稻阳性单株;CK.转基因水稻阴性单株(没有该检测基因)。

Fig.3 Identification of positive transgenic rice lines by PCR M.DL2000 bp DNA Marker;1-9.Some positive individuals of transgenic rice; CK.Negative individuals of transgenic rice(no gene for this test).

图4 转基因水稻中TaGAMyb-B基因的相对表达量

Fig.4 The relative expression levels of TaGAMyb-B in transgenic rice

图5 不同胁迫处理下转基因水稻中TaGAMyb-B基因的表达变化

Fig.5 Expression changes of TaGAMyb-B in transgenic rice with during different stress treatments

图6 转基因水稻表型统计分析不同小写字母表示差异显著,P<0.05。图8同。

Fig.6 Statistic and analysis of phenotype in transgenic rice Different lowercase letters indicate significant different,P<0.05.The same as Fig.8.

图7 转基因水稻第二节间横切面

Fig.7 Transverse sections of second stem section in transgenic rice

图8 转基因水稻第二节间横切面厚壁组织平均厚度

Fig.8 Average thickness of sclerenchyma in transverse sections of second stem section in transgenic rice

图9 转基因水稻第二节间纵切面

Fig.9 Longitudinal sections of second stem section in transgenic rice

图10 转基因水稻第二节间纵切面细胞平均长度不同大写字母表示差异极显著,P<0.01。

Fig.10 Average length of cell in longitudinal sections of second stem section in transgenic rice Different capital letters indicate extremely significant,P<0.01.

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